Sublethal effects of beta-cypermethrin on the bird cherry-oat aphid Rhopalosiphum padi (Hemiptera: Aphididae)

Rhopalosiphum padi is a sap-sucking aphid and an important pest of wheat that causes considerable yield loss. Beta-cypermethrin, a synthetic pyrethroid pesticide, has a broad insecticide spectrum and is considered effective for aphid control, while its residual concentrations may have sublethal effects on R. padi. Here, the sublethal effects of beta-cypermethrin on R. padi were conducted under laboratory conditions. The acute toxicity test showed that LC₁₀, LC₂₀, and LC₂₅ of beta-cypermethrin to R. padi adults were 0.003, 0.031 and 0.079 mg L⁻¹, respectively. The pre-adult survival rate was significantly reduced by all three concentrations. LC₂₀ significantly extended the development duration of 1st instar nymphs, pre-oviposition period, and oviposition period of R. padi. The adult longevity was also reduced by LC₂₅. However, the fecundity did not differ between the beta-cypermethrin treatment and control. For life table parameters, both the finite rate (λ) and intrinsic rate of increase (r) decreased at LC₁₀ and LC₂₀, as well as the net reproductive rate (R₀) reduced at LC₁₀ and LC₂₅, while mean generation time (T) increased at LC₂₀. Thus, at the concentrations of beta-cypermethrin tested here, there were negative impacts on R. padi fitness by decreased pre-adult survival rate, λ, r, and R₀, and delayed the development of some stages and increased T.     

Resistance of wheat lines to Rhopalosiphum padi (Hemiptera: Aphididae) under laboratory conditions

Bird cherry-oat aphid, Rhopalosiphum padi (L.), a polyphagous species with a nearly worldwide distribution, is an important pest of wheat as well as the main vector of barley yellow dwarf virus. We evaluated the resistance categories of eight wheat lines including antibiosis, antixenosis, and tolerance against R. padi under laboratory conditions. The wheat lines tested were ERWYT 88-8, ERWYT 87-6, and ERWYT 87-4 (resistant); ERWYT 87-1, ERWYT 87-20, and ERWYT 88-11 (susceptible); ERWYT 88-12 and ERWYT 88-13 (intermediate). In the antibiosis experiment, R. padi produced fewest progeny on ERWYT 88-8, ERWYT 87-6, and ERWYT 87-4 in reproduction period. In the antixenosis test, R. padi performed best on ERWYT 87-1, ERWYT 87-20, and ERWYT 88-11. Fewer apterous aphids selected ERWYT 88-8, ERWYT 87-4, and ERWYT 87-6 lines indicating antixenosis of these lines to R. padi. In tolerance experiments, however growth parameters differed between treated and untreated seedlings of wheat lines with 10 aphids per day infestation during 21-d period, but not among eight wheat lines. The plant resistance index values were greatest for ERWYT 88-8 (9.71), followed by ERWYT 87-4 (7.04) and ERWYT 87-6 (4.76). ERWYT 88-8, ERWYT 87-6, and ERWYT 87-4 may be important sources of R. padi resistance for small grain breeding and integrated pest management programs.     

禾谷缢管蚜鞣化激素基因克隆及功能分析

【目的】探讨禾谷缢管蚜 Rhopalosiphum padi （Linnaeus）鞣化激素基因的发育表达模式及功能。【方法】采用转录组测序得到禾谷缢管蚜鞣化激素基因bursicon-α 和 bursicon-β cDNA序列。通过实时荧光定量PCR方法,分析该基因的发育表达模式。利用RNA干扰（RNA interference, RNAi）介导 bursicon-α 和bursicon-β 沉默,分析鞣化激素的功能。【结果】序列分析结果显示,禾谷缢管蚜鞣化激素α亚基基因（bursicon-α）cDNA序列开放阅读框为480 bp,编码159个氨基酸残基;β亚基基因（bursicon-β）cDNA序列开放阅读框为417 bp,编码138个氨基酸残基。时序表达分析表明,鞣化激素两个亚基基因在禾谷缢管蚜整个发育期均有表达,以1龄若蚜期表达量最高;成蚜有翅个体中表达量显著高于无翅个体。RNAi介导的 bursicon-α 和 bursicon-β 沉默均能显著抑制禾谷缢管蚜成蚜表皮的黑化。【结论】研究结果表明,鞣化激素在禾谷缢管蚜体壁黑化中发挥着重要作用。该结果为进一步研究鞣化激素在蚜虫生长发育过程中的生理功能提供了基础资料。     

Ultrastructure des cornicules de Rhopalosiphum padi L. (Homoptera : Aphididae)

The cornicles, exclusive structures of aphids (Homoptera : Aphididae), are distinct secretory organs and are comprised of: (1) a group of cornicle secretory cells having spatial and temporal maturation, enclosed within a sac, which has a structure identical to that of the basal membrane of the epidermis; (2) cuticular structures, valves permitting the emission of the secretion. The maturation of the secretory cells follows a linear sequence from the basal area to the apex of the cornicle. When their differentiation is finished, these cells are characterized by the presence of an important central inclusion of lipid and numerous vacuoles containing a volatile substance. In response to an external stimulus, the cornicles extrude a droplet of secretions having a defensive function, either disturbing the predators or inducing the dispersion of the aphids, or both.     

A novel theta-class glutathione S-transferase gene in Rhopalosiphum padi (L.) (Hemiptera: Aphididae): Identification,recombinant protein expression and function analyses of RpGSTT1

Glutathione S-transferases (GSTs) play a major role in the detoxification of insecticides and the protection against oxidative damage caused by ROS (reactive oxygen species). The bird cherry-oat aphid, Rhopalosiphum padi (L.), is a major crop pest worldwide. Chemical insecticides are currently used to control aphids, but insecticide resistance is an evolutionary issue for aphid control. In this research, we identified and cloned the full-length cDNA of a theta-class GST gene (RpGSTT1) from R. padi, which contains 702 bp open reading frame encoding 233 amino acids. RpGSTT1 was clustered together with theta classe of insect GSTs in the phylogenetic tree. Effect of insecticide exposure on the GST enzyme activity was investigated in vivo. After affinity purification, the recombinant RpGSTT1 protein was able to catalyze the conjugation of 1-chloro-2, 4-dinitrobenzene (CDNB). A disc diffusion assay showed that the RpGSTT1 can increase resistance to cumene hydroperoxide-induced oxidative stress. In addition, the purified recombinant RpGSTT1 protected super-coiled DNA from oxidative damage. By quantitative PCR, The relative expression level of RpGSTT1 in the aphid was highly up-regulated after 12 h exposure to β-cypermethrin, isoprocarb, malathion and sulfoxaflor, respectively. To our knowledge, RpGSTT1 is the first theta class of GST gene reported in bird cherry-oat aphid. Our findings indicate that RpGSTT1 may be involved in insecticide detoxification in R. padi.     

Feeding behaviour of the aphid Rhopalosiphum padi (Hemiptera: Aphididae) on nitrogen and water-stressed barley (Hordeum vulgare) seedlings

Electrical penetration graphs (EPGs) were used to examine the probing behaviour of adult apterous Rhopalosiphum padi (Linnaeus) on barley seedlings grown under conditions of nitrogen or water stress. Aphids took significantly longer to reach and ingest from sieve elements of nitrogen-deficient seedlings than from nitrogen-sufficient seedlings but there were no such differences between water-stressed or well-watered seedlings. On both nitrogen and water-stressed seedlings the average length of each individual period of salivation into the sieve element was significantly greater compared with their respective unstressed controls.     

Induced proteolysis within the bird cherry leaves evoked by Rhopalosiphum padi L. (Hemiptera, Aphidoidea)

The objectives of this study were to elucidate the impact of bird cherry-oat aphid (Rhopalosiphum padi L.) feeding on functioning of the proteolytic machinery in bird cherry leaves. Biochemical analyses proved that R. padi feeding in tissues of primary host stimulated activity of the two major fractions of proteinases (extracted at the optimal pH values: 5.0 and 7.0). Additionally, it has been demonstrated that aphids' feeding on bird cherry led to a decline in levels of albumins and globulins (main protein fractions in P. padus leaves). The opposite tendency, regarding the amounts of these protein fractions was ascertained at the phase of disappearance of R. padi population on tested shoots. Furthermore, it is reported that an increase in activity of the analysed enzymes and a decline in the content of tested protein fractions, were proportional to density of aphid individuals developing on P. padus side shoots. It is hypothesized that long-term R. padi feeding may lead to intensifying the catabolic processing of proteins by the activated proteolytic machinery in bird cherry leaves. The multi-level biological functions of endogenous plant proteinases and their significance in triggering the defense reactions in aphid-infested plant tissues are discussed.     

Transgenic Bt maize and Rhopalosiphum padi (Hom., Aphididae) performance

Abstract.  1. The population abundance and age structure of Rhopalosiphum padi , one of the most common maize aphid species, on transgenic Bt (expressing the Cry1Ab protein) and non-Bt isogenic maize was studied in commercial plots during three crop seasons.
2. A higher density of aphids, particularly alates and young nymphs, occurred in Bt plots at very young maize development stages, corresponding to the settlement period, in the 3 years studied. Possible causes are discussed. After this period, there were no differences between Bt and non-Bt maize.
3. Mortality, development, and reproduction of the offspring of alate forms of R. padi and the offspring of different generations of apterous forms fed with Bt maize were evaluated in the laboratory under controlled conditions.
4. The developmental and pre-reproductive times of the offspring of the first generation of alatae were shorter and the intrinsic rate of natural increase ( r _m ) higher when aphids fed on Bt maize. The opposite occurs with the offspring of the first generation of apterous mothers, which have lower nymphal and adult mortality, shorter developmental and pre-reproductive times, a higher effective fecundity rate, and greater r _m , when fed on non-Bt maize. The differences in aphid development on the two cultivars may be linked to changes in host-plant quality due to pleiotropic effects of the genetic modification.
5. No differences on aphid mortality, developmental and pre-reproductive times, fecundity, and r _m were found between the offspring of apterous aphids maintained on Bt or non-Bt maize for several generations.     

The effect of natural enemies on the spread of barley yellow dwarf virus (BYDV) by Rhopalosiphum padi (Hemiptera: Aphididae)

The effects of two natural aphid enemies, adult Coccinella septempunctata Linneaus, a predator, and Aphidius rhopalosiphi de Stefani Perez, a parasitoid, on spread of barley yellow dwarf virus (BYDV) transmitted by the bird cherry-oat aphid, Rhopalosiphum padi (Linnaeus) were studied under laboratory conditions. Predators or parasitoids were introduced to trays of durum wheat seedlings and the patterns of virus infection were observed after two, seven and 14 days of exposure. More plants were infected with BYDV in control trays without A. rhopalosiphi than in trays with the parasitoid present, both seven and 14 days after the introduction of parasitoids. Patterns of virus infection were found to be similar over time in trays with a parasitoid present and in control trays. More plants were infected in trays with C. septempunctata present than in control trays, both two and seven days after the introduction of the coccinellid. The spread of virus infections progressed differently over time for the two treatments (predator and parasitoid), differences between treatments being most marked after two days and seven days, when more plants exposed to predators but fewer exposed to parasitoids were infected with BYDV compared to their respective controls. However, by the 14th day 88% of all plants were infected and there was no significant difference between the two treatments. The role of natural enemies in spread of BYDV is discussed.     

Diuraphis noxia and Rhopalosiphum padi (Hemiptera: Aphididae) interactions and their injury on resistant and susceptible cereal seedlings

Interspecific interactions between the symptomatic (chlorosis-eliciting) Russian wheat aphid, Diuraphis noxia (Mordvilko), and the asymptomatic (nonchlorosis-eliciting) bird cherry-oat aphid, Rhopalosiphum padi (L.), on four cereal genotypes were examined by simultaneous infestations. Four cereals (i.e., Diuraphis noxia-susceptible 'Arapahoe' wheat and 'Morex' barley, and D. noxia-resistant 'Halt' wheat and 'Border' oat) and four infestations (i.e., control, D. noxia, R. padi, and D. noxia/R. padi) were used in the research. Whereas D. noxia biomass confirmed D. noxia resistance among the cereals, R. padi biomass indicated that the D. noxia-resistant cereals did not confer R. padi resistance. D. noxia biomass was significantly lower in D. noxia/R. padi infestation than that in D. noxia infestation on all cereals, except Border oat, which indicated an antagonistic effect of R. padi on D. noxia. All aphid infestations caused a significant plant biomass reduction in comparison with the control. In comparison with D. noxia infestation, D. noxia/R. padi caused a significant plant biomass reduction on all cereals, except Morex barley. Although D. noxia biomass in D. noxia/R. padi infestation was significantly less than that in D. noxia infestation, leaf chlorophyll reduction was the same between D. noxia/R. padi and D. noxia infestations, which suggested that the asymptomatic R. padi enhanced the D. noxia-elicited leaf chlorophyll loss. The regression between chlorophyll content and aphid biomass indicated that the asymptomatic R. padi in the D. noxia/R. padi infestation enhanced chlorophyll loss, but interspecific aphid interaction on plant biomass varied among the cereals.     

Distribution of Rhopalosiphum padi (Hornoptera: Aphididae) on spring barley plants

The distribution of Rhopalosiphum padi on spring barley plants in the field and the laboratory is described. Aphids fed mainly on the lower parts of the plants in both the field and the laboratory. A relatively large proportion of the field population was found beneath the soil surface early in the season. In the laboratory, aphids preferred the stem base and auricles of young and old plants respectively. The highest soluble nitrogen concentration was found in the stem base of young plants. The aphids avoided temperatures above 25°C in a temperature gradient (20°C-40°C). The soil surface temperature in a barley stand can reach lethal levels for aphids during clear days early in the season. It is concluded that the preference of R. padi for the lower part of young spring barley plants is mainly a consequence of the stem base being the most nutritious part of the plant and that a harsh microclimate possibly forces the aphids beneath the soil surface. This tendency to feed near or below soil level must be taken into account when assessing field densities of R. padi.     

Distribution and molecular composition of heterochromatin in the holocentric chromosomes of the aphid Rhopalosiphum padi (Hemiptera: Aphididae)

In order to study the structure of holocentric chromosomes in aphids, the localization and the composition of Rhopalosiphum padi heterochromatin and rDNA genes have been evaluated at cytogenetic and molecular level. In particular, heterochromatin resulted located on all the chromosomes both in intercalary and telomeric positions. Moreover, enzymatic digestion of R. padi genome put in evidence a DraI satellite DNA which has been isolated, cloned and sequenced. FISH experiments showed that this satellite DNA clusters in an intercalary C-positive band on the two X chromosomes.     

禾谷缢管蚜复眼外部形态的扫描电镜观察

【目的】通过比较禾谷缢管蚜Rhopalosiphum padi两种生活史(全周期型和不全周期型)及5种蚜型(有翅孤雌蚜、无翅孤雌蚜、雄蚜、雌性蚜和产雌性母)成蚜复眼外部形态,分析了其视觉能力差异,为进一步探索禾谷缢管蚜生殖转换及寄主搜寻机制提供参考。【方法】利用扫描电镜技术,观察全周期型和不全周期型禾谷缢管蚜无翅孤雌蚜及其低温短日照(12℃, 光周期8L∶16D)诱导条件下全周期型所产5种蚜型成蚜的复眼外部形态。【结果】禾谷缢管蚜成蚜复眼着生于头部触角基部,呈卵圆形,小眼间无感觉毛。全周期型和不全周期型禾谷缢管蚜无翅孤雌蚜的复眼大小(长轴直径及短轴直径)、复眼弧高、小眼数量和小眼直径均没有显著差异。而这5个复眼指标在低温短日照诱导全周期型禾谷缢管蚜产生的5种蚜型的成蚜中均存在显著性差异,其中,雄蚜、有翅孤雌蚜和产雌性母的复眼区域较大,小眼数量较多;雌性蚜的复眼区域最小,小眼数量也最少。【结论】综合以上指标结果,推测5种蚜型的视觉能力由大到小是：雄蚜＞有翅孤雌蚜＞产雌性母＞无翅孤雌蚜＞雌性蚜。复眼和小眼的这些差异可能与禾谷缢管蚜生殖转换及寄主搜寻存在一定的联系。     

禾谷缢管蚜阳离子通道基因RSC1的克隆、分子特性及诱导表达分析

【目的】SC1通道(sodium channel 1)是昆虫体内一种重要的离子通道,被认为是一种开发新型杀虫剂的神经靶标。本研究拟克隆禾谷缢管蚜Rhopalosiphum padi的SC1通道基因,并初步分析其生理功能及其与SC1类通道、电压门控钠离子通道、电压门控钙离子通道的进化关系。【方法】采用RT-PCR技术,克隆了禾谷缢管蚜SC1基因完整的开放阅读框;利用实时荧光定量PCR技术,分析禾谷缢管蚜成蚜在不同浓度的高效氯氟氰菊酯诱导下SC1基因表达变化。【结果】获得了禾谷缢管蚜SC1基因(命名为RSC1)完整的开放阅读框(Gen Bank登录号为KU640190),其长度6 687bp,编码2 228个氨基酸。RSC1具有SC1通道的结构特征,有一个不同于电压门控钠离子通道和电压门控钙离子通道的特殊DEEA模体(motif)。系统进化分析结果显示,RSC1与电压门控钠离子通道组成一个进化枝,电压门控钙离子通道组成另外一个进化枝,SC1与电压门控钠离子通道在进化上有更近的起源关系。实时荧光定量PCR分析结果表明,LC15,LC35和LC503种剂量的高效氯氟氰菊酯处理6 h后,禾谷缢管蚜RSC1基因表达量相对于清水对照显著下调,表达量分别为对照的0.57,0.82和0.78倍;3种剂量的高效氯氟氰菊酯处理24 h后,禾谷缢管蚜RSC1基因表达量分别为对照的2.19,1.33和1.19倍,其中LC15(0.1484 mg/L)胁迫下RSC1基因的表达量显著上调。【结论】SC1类通道与电压门控钠离子通道在进化起源上有更近的关系。RSC1通道可能是高效氯氟氰菊酯的次级靶标。由于RSC1和其同源基因只存在于节肢动物中,脊椎动物尚未发现该类基因,因此这类通道可能作为开发新型杀虫剂的神经靶标。     

Assessment of potential impacts due to unintentionally released Bt maize plants on non-target aphid Rhopalosiphum padi (Hemiptera: Aphididae)

Genetically modified maize crops expressing Bacillus thuringiensis (Bt) toxins (Bt maize) are increasingly cultivated worldwide, and large amounts of Bt maize have been imported to Korea. Before evaluating the environmental impacts of Bt maize of unknown origin on non-target insects, crystal (Cry) protein types in the imported Bt maize plants were identified. Because Cry1F was found in the tested Bt maize plants, Rhopalosiphum padi, a non-lepidopteran species, was selected as the non-target insect species. Additionally, a widely cultivated domestic maize strain was selected as an alternative control. No difference in survival rate, alata vivipara production, or host preference was observed between R. padi fed on the Bt maize and the control non-Bt maize, indicating that Bt maize plants had no sub-chronic adverse effects on R. padi. The average number of nymphs from Bt maize-fed aphids was 1.73-fold higher than that of non-Bt maize-fed aphids, implying that R. padi population density can increase after several generations in Bt maize fields. An enzyme-linked immunosorbent assay revealed that Cry1F toxin concentrations increased gradually in the body of R. padi when they were fed Bt maize, but that all ingested Cry toxins were excreted within 10 days after Bt-fed aphids were transferred to non-Bt maize, suggesting little possibility of Cry toxin exposure via R. padi to the endoparasitoids. However, the possibility still remains that Cry toxins can be transferred to predatory insects in higher trophic levels if they consume Bt maize-fed aphids.     

禾谷缢管蚜ABC转运蛋白基因的克隆、分子特性及表达分析

【目的】ABC（ATP-binding cassette）转运蛋白是一类重要的跨膜蛋白超家族,某些ABC转运蛋白基因在一些害虫的抗药性品系中表达显著提高。本研究旨在克隆禾谷缢管蚜 Rhopalosiphum padi ABC转运蛋白基因RhpaABCG9,RhpaABCG20和RhpaABCG23 cDNA全序列,分析这3个基因在该虫不同发育阶段和不同抗药性品系中的表达模式,为阐明ABC转运蛋白在禾谷缢管蚜抗药性中的作用和其他生理功能,以及深入分析该虫抗药性机理奠定基础。【方法】采用RT-PCR与RACE技术,克隆了基因cDNA全序列;利用实时荧光定量PCR技术,研究这3个基因在禾谷缢管蚜不同生长发育阶段和不同抗药性品系中的表达变化。【结果】获得了RhpaABCG9,RhpaABCG20和RhpaABCG23 3个基因cDNA全序列,其开放阅读框长度分别为2 103,2 436 和2 082 bp,分别编码700, 811和693个氨基酸。结构分析表明,3个蛋白均具有ABC转运蛋白家族典型的结构特征;系统进化分析结果显示,3个蛋白氨基酸序列分别与豌豆蚜Acyrthosiphon pisum各对应氨基酸的序列一致性最高。实时荧光定量PCR分析结果表明,这3个基因在禾谷缢管蚜不同发育时期均不同程度表达。RhpaABCG20在各个发育时期的表达变化差异不显著;RhpaABCG9表达量在4龄若蚜最高,在1龄若蚜最低;RhpaABCG23 表达量在3龄若蚜最高,1龄若蚜最低,其他阶段差异不显著。禾谷缢管蚜异丙威抗性品系中,RhpaABCG20表达量显著高于敏感品系,而RhpaABCG9和RhpaABCG23表达量均低于敏感品系,但差异不显著;禾谷缢管蚜吡虫啉抗性品系中,RhpaABCG20和RhpaABCG23表达量显著高于敏感品系,而RhpaABCG9表达上调不显著。【结论】RhpaABCG9,RhpaABCG20和RhpaABCG23基因可能参与禾谷缢管蚜体内农药的运输,并与禾谷缢管蚜的抗药性具有一定的关系。本研究结果为进一步深入分析禾谷缢管蚜的抗药性机制,以及该虫的抗药性治理与综合防治奠定了基础。     

Odour stimuli affecting autumn migration of Rhopalosiphum padi (L.) (Hemipter: ornoptera)

R. padi gynoparae settling in the canopy of the winter host, Prunus padus, tended to form aggregations. Differences in the degree of exposure to wind, sun, etc., did not satisfactorily explain the aggregated pattern observed. In laboratory experiments, aphids did not distinguish between leaves taken from nearby aggregations and leaves taken far away from aggregations. The results of laboratory olfactometer tests and field trapping studies indicate that R. padi gynoparae produce an aggregation pheromone. Thus the aggregated settling pattern of gynoparae of R. padi can best be explained as a response to an aggregation pheromone.     

Antibiosis and Non-preference of Sitobion avenae (F.) (Hemiptera: Aphididae) on Leaves and Ears of Commercial Cultivars of Wheat (Triticum aestivum)

Little is known on the resistance of wheat cultivars to Sitobion avenae (F.) in Brazil. The goal of this work was to assess the behavior and biology of S. avenae on four commercial wheat cultivars to verify the existence of resistance by antibiosis in leaves and ears and non-preference in the ears. The smallest net fecundity rates of S. avenae in wheat leaves have been found in the cultivars Embrapa 22 and BRS264, which did not differ between themselves. The intrinsic rate of increase of S. avenae was smaller in leaves of Embrapa 22 than in cultivars BRS254 and BRS Timbaúva. The smallest net fecundity rates of S. avenae in wheat ears were observed in the cultivars BRS254 and Embrapa 22. The intrinsic rate of increase of the aphid in the ear of cultivar Embrapa 22 was smaller than in BRS Timbaúva and BRS264, but did not differ from BRS254. The organ of the wheat plant in which the aphid was reared influences antibiosis resistance, but the cultivar BRS Timbaúva was considered susceptible and Embrapa 22 resistant to S. avenae in both plant organs tested. Ears of wheat cultivars tested did not show differences in the mechanism of resistance by non-preference to S. avenae.